Studies on arylsulphatases in the barnacle Balanus eburneus

Arylsulphatases (ASases) have been identified in the mantle tissues of the barnacle Balanus eburneus Gould. This is the first report of these enzymes in the Cirripedia. Using p-nitrocatechol sulphate (NCS) as a substrate, the optimum pH for activity was found to be 5.6 in 0.5 M-acetate buffer, and substrate concentration of 12 mM. The activity of the enzyme was strongly inhibited by phosphate, sulphite and sulphate and not by cyanide, indicating the presence of Type II ASase. Biochemical and electrophoretic studies revealed the coexistence of at least two distinct Type II ASases in the mantle tissues. These are probably similar to the ASase-A and ASase-B reported for vertebrate tissues. The presence of multiple molecular forms of ASase capable of hydrolyzing potassium 6-benzoyl-2-naphthyl sulphate was demonstrated by polyacrylamide gel disc-electrophoresis. The activity patterns of ASases in the mantle tissues were studied in relation to the molt cycle. The activity of ASase-B followed a cyclic pattern which correlated with the molt stages, reaching a maximum in the postmolt. The activity of ASase-A remained essentially the same during the entire molt cycle. Analysis of the activities after dialysis indicates a change in the activity of ASase-B into a non-fluctuating pattern. The enzyme was found in all stages of development, with a ninefold increase in activity in adult forms. The fact that a greater activity was found in the nondigestive organs and that there is an increase in the activity of ASase in tissues of unfed (starved) specimens indicates that the enzyme does not function in the digestive processes, as suggested for other animals by certain investigators. The evidence in this study implicates a major role for dialyzable inhibitory factor(s) as the mechanism involved in the regulation of the activity of ASase-B during the molt cycle. The possible relationship of endogenous phosphate and/or neurosecretory substance(s) with the dialyzable inhibitory factor(s) is discussed. A discussion speculating on the relationship of ASase activity to the cyclic formation and hardening of the exoskeleton and adhesive substance(s) is also presented.     

Energy flow and population dynamics of the barnacle Balanus glandula

The energetics and population dynamics of a barnacle (Balanus glandula Darwin) population in British Columbia, Canada, were studied. Consumption, energy flow, production and mortality were 6844.6, 6667.0, 2896.5 and 2522.8 Kcal m^-2 year^-1, respectively. These energy flow and production values are among the highest for animal populations reported, and therefore strongly suggest the functional importance of E. glandula in littoral systems. The young age groups of the first-year settlements were most important in contributing to the energy flow, production and reproduction of the entire population. Most of the assimilated energy in the older age groups was used in respiration.     

“Copper” granules in the barnacle Balanus balanoides

Balanus balanoides (L.) collected from an area with high heavy-metal run-off contained two different types of granule within the parenchyma cells of the prosoma. X-ray microprobe analysis shows one to be the familiar zinc granule made up of concentric layers and giving major peaks for phosphorus and zinc, and the other to be homogeneous and giving peaks for sulphur and copper. This latter granule is designated the copper granule. Whilst zinc granules are known to be composed of inorganic phosphate, various tests on copper granules in sections and in a granule-rich pellet have shown that the copper is probably complexed with organic matter. The tests also demonstrated the relatively, insoluble (inert) nature of these granules. Although zinc and copper granules were present together in the prosoma, atomic absorption analyses of whole bodies (prosoma+thorax) have shown the level of zinc (50.28 g/mg dry weight) to be much higher than that of copper (3.75 g/mg dry weight).     

Zinc phosphate granules in tissue surrounding the midgut of the barnacle Balanus balanoides

The chemical composition of inorganic granules found in parenchyma cells surrounding the midgut of adult Balanus balanoides (L.) was determined. X-ray microanalysis indicated the presence of magnesium, phosphorus, potassium, calcium, iron and zinc in the granules. A quantitative analysis of granule-rich pellets isolated from the midguts of adult barnacles showed that the granules were composed mainly of zinc phosphate, the remaining elements being minor constituents only.     

Acute toxicity and sublethal behavioral effects of copper on barnacle nauplii (Balanus improvisus)

This study documents the effects of short-term (24h) sublethal copper exposures on undirected swimming activity and photobehavior of Balanus improvisus stage II nauplii. All Cu treatments were static, with temperature and salinity conditions at 20°C and 15 or 30. The 24h LC 50 estimate for Cu is 88 ppb at 15 and >200 ppb at 30. Sub-lethal Cu concentrations cause reductions in swimming speed, which decrease progressively with increasing Cu dose. At 50 ppb Cu, this was significant primarily at light intensities below the phototactic threshold. At higher Cu concentrations, significant reductions in mean linear velocity occurred at most light intensities tested. At 30, 50 and 100 ppb Cu also reduce the positive phototactic response and 150 ppb Cu causes reversal of phototaxis at optimal intensities. Photokinesis is reduced at 100 ppb Cu and disappears at 150 ppb Cu. At 15, the behavioral effects of 50 ppb Cu resemble those occurring with 150 ppb Cu at 30. Swimming speed and photobehavior show promise as sensitive behavioral indicators of copper toxicity. Additional research is required to determine if these responses apply to a broad range of pollutants and to other planktonic organisms. There is also a need to further evaluate the significance of these behavioral effects ecologically.Contribution No. 181 from the EPA Environmental Research Laboratory, Narragansett, RI 02882, USA     

Characterization of barnacle (Balanus eburneus and B. cenatus ) adhesive proteins

Cements from two species of barnacles, Balanuseburneus Gould and Balanus crenatus Bruguiére, were analyzed to identify the number and nature of proteins present. B. crenatus cement was composed mainly of a small peptide cross-linked into aggregates ranging in size from ˜ 3 kD to 40–50 kD. These aggregates could be reduced with 2-mercaptoethanol (2-ME) in the presence of sodium dodecyl sulfate (SDS) but only after incubation at 40 °C for 12 to 24 h and only when the cement was recently formed. Reductive alkylation of cysteine residues with 4-vinylpyridine produced a water-soluble peptide of less than 5 kD. By comparison, the cement of B. eburneus could be dissolved only partially in SDS and 2-ME when heated at 100 °C for 10 min. Five major proteins were identified by SDS-PAGE: 7, 22, 36 and 58 kD bands for which N-terminal sequence and amino acid compositions are presented; and a 52 kD band for which sequence data are given. A minor protein band of ˜ 80 kD has the same N-terminus as the 36 kD band. CNBr digests of individual proteins produced peptides for which sequence and composition data are also presented. The study was conducted during 1993 to 1995. In general, the proteins identified from B. crenatus cement are similar to those characterized from B. eburneus, and they are different in composition and sequence than those previously reported from Mytilus edulis. Received: 7 August 1996 / Accepted: 8 October 1996     

Influence of substratum heterogeneity and settled barnacle density on the settlement of cypris larvae

On the Atlantic coast of Canada, Semibalanus balanoides (L.) is widely distributed in the mid-intertidal zone, whereas in the Gulf of St. Lawrence, this species is mostly limited to crevices. We tested the hypothesis of regional differences in microhabitat selection by barnacle larvae at settlement in 1984 and 1985 at St. Andrews, New Brunswick, Canada. Since larvae settle in microhabitats already colonized by adults, the relative influence of settled barnacle density and of different scales of substratum heterogeneity on settlement were evaluated experimentally at Capucins, Québec, (Gulf of St. Lawrence) and at St. Andrews, New Brunswick (Atlantic coast). On a large scale (>10 cm deep crevices) of heterogeneity, results show that, in the Gulf, cypris larvae settled nearly exclusively (93%) in natural crevices rather than on adjacent horizontal surfaces. On the Atlantic coast, settlement was more important outside than inside of crevices, when the substrata were either natural or artificial. This result is unique and contrasts sharply with all known reports on barnacle settlement in relation to surface contour. The influence of barnacle density on settlement was greater than that of large scale heterogeneity. On a small scale (<1.5 cm deep cracks), the presence of conspecifics had a stronger effect on settlement than heterogeneity in both regions. Field observations showed a relationship between larval settlement density and percentage of adult cover. Settlement increased up to 22 or 30% (Gulf and Atlantic coast) of adult cover and decreased afterwards. The results confirm the hypothesis of larval selection for cryptic habitats in the Gulf and the opposite behaviour (preferences for horizontal surfaces) on the Atlantic coast. This microhabitat selection is apparent at large scales of heterogeneity, whereas at small scales, the presence of conspecifics is the predominant factor.Contribution to the programme of GIROQ (Groupe interuniversitaire de recherches océanographiques du Québec)     

Noncovalent bonds are key mechanisms for the cohesion of barnacle (Balanus crenatus) adhesive proteins

Cement of the barnacle species, Balanus crenatus, was isolated and separated by SDS-PAGE. Reductive and nonreductive conditions yielded the same result of six major proteins (in the range up to 100 kDa) occurring as two triplets. A third triplet of lower molecular mass was faintly visible. The presence of a reductive agent had only little impact on the solubility of B. crenatus cement. The strong effect of the denaturing conditions (the presence of SDS and the incubation at elevated temperature) on the solubility of the barnacle cement and the absence of disulfide signals in Raman spectra suggest a complex structure of the barnacle cement noncovalently bound. It can be speculated that this protein complex in its quaternary structure is formed by small subunits and is equivalent to the globular cement structures previously detected.     

Initial contact,exploration and attachment of barnacle (Balanus amphitrite) cyprids settling in flow

Settlement responses of barnacle (Balanus amphitrite) cyprids to boundary-layer flows were examined in laboratory flume-experiments. The leading-edge configuration of flat plates was altered in order to manipulate flows without changing surface topography or freestream velocity. Settlement along the plates correlated strongly with downstream gradients in shear stress. Analyses of video images taken during the experiments indicate that cyprids first contact plates in regions where plate-ward advection is high, and subsequent exploratory movement along the plate is oriented with flow direction at the plate surface. After exploration, cyprids reject a surface more frequently in a fast flow (10 cm s^–1 freestream velocity) than in a slow flow (5 cm s^–1), but rejection occurs in shear stresses well below the threshold that would prevent attachment and exploration. A higher rejection rate does not result in lower settlement, however, since contact rate is higher in fast than slow flows. The movement of cyprids in flow thus appears to be a passive transport process during the initial contact stage of settlement, but an active behavioral response to flow direction and shear stress during later stages of exploration and attachment.     

The histology,histochemistry and ultrastructure of the cement apparatus of three adult sessile barnacles,Elminius modestus,Balanus balanoides and Balanus hameri

The histology, histochemistry and ultrastructure of the cement gland cells of 3 species of sessile barnacles are described. Cells of both Elminius modestus Darwin and Balanus balanoides (L.) (membranous bases) are similar histologically and ultrastructurally, and within the cells there are 2 distinct regions. One is the site of secretion accumulation, the other the site of secretion synthesis. In Balanus hameri Ascanius (calcareous base) the cells are different in that the cytoplasm is more homogeneous. In all 3 species protein is the final secretion product (cement) of the cement cells. The mode of discharge of secretion from the cells differs between the membrane based and calcareous based barnacles. In the former, there is a series of collecting canals within the cytoplasm of the cement cells (intracellular canals) which join with the larger extracellular cement ducts. Secretion passes into the intracellular canals and is moved along to the larger cement ducts, which have an inner chitin lining. In Balanus hameri (calcareous base) there are no intracellular collecting canals. Secretions are thought to pass from the cement cells into the cement duct cells direct. The cement ducts in this species are not chitin lined.     

Temperature and salinity interactively impact early juvenile development: a bottleneck in barnacle ontogeny

When juvenile mortality or juvenile growth is impacted by temperature and salinity, these factors have a substantial effect on recruitment success and population dynamics in benthic ecosystems. Using freshly settled cyprids of Amphibalanus improvisus, we investigated the combined effects of temperature (12, 20 and 28 °C) and salinity (5, 15 and 30 psu) on early juvenile stage performance. Mortality as well as size (basal diameter, dry weight, and ash-free dry weight) was monitored for a period of 40 days. Mortality was high (42–63 %) during the first week following attachment, regardless of the temperature and salinity treatments. Subsequently, mortality and size were interactively influenced by temperature and salinity. Highest mortality and lowest size of juveniles occurred at lowest temperature (12 °C) and salinity (5 psu). Apparently, low temperature (12 °C) narrowed the barnacles’ salinity tolerance. Juvenile barnacles constructed more shell material compared to body mass at high temperature and high salinity, while a reverse situation was observed at low temperature and low salinity. Our results demonstrate that environmental changes can directly and/or indirectly alter patterns of survivorship and size. Warming and desalination as predicted for the Baltic Sea in the course of climate change may, however, act antagonistically and compensate each other’s isolated effect on barnacles.     

Development of the gooseneck barnacle Pollicipes polymerus (Cirripedia: Lepadomorpha): Fertilization through settlement

Pollicipes polymerus (Sowerby, 1833) is a common barnacle in rocky intertidal zones of the West Coast of North America whose development has been previously undescribed in detail. In the present study, certain aspects of the development of P. polymerus are recorded from fertilization through settlement. In vitro fertilization has been accomplished using already-implanted sperm plus ovulating eggs and oviducal gland fluid. The eggs are small (about 100 μ diameter at fertilization; 90 x 140 μ at first cleavage) and somewhat yolky. Embryos obtained from brooding adults and from in vitro fertilization were reared in glass dishes until hatching. Cleavage was total, asynchronous and unequal. Division of the D macromere lagged behind division of the other blastomeres, and gastrulation occurred by epiboly so that blastoderm surrounded the macromeres. The nauplii hatched an average of 25.4 days (range was 20.3 to 30.1 days) after fertilization, and molted within 1 day to Stage II nauplii. From Stages II through VI the nauplii required food. As they grew from stage to stage, the number of setae per appendage increased. Nauplii have feathery and hispid-type setae, previously described only in Chthamalus aestuarii. In contrast to other pedunculate larvae which have been described, P. polymerus nauplii are small and have specialized feathery setae. Cypris-larva settlement was stimulated only when healthy adult peduncles were available, thus showing an affinity for its own species.     

Effects of crowding and wave exposure on penis morphology of the acorn barnacle, Semibalanus balanoides

Wave action and low population density can strongly reduce the ability of sessile acorn barnacles to find mates and copulate. For Semibalanus balanoides, penis morphology varies with wave exposure and with characteristics of the mating neighborhood. Field experiments were conducted at five intertidal sites on Long Island, New York, USA from July to December 2005 to determine how wave exposure and aggregation structure influence the length, diameter, mass, and number of annulated folds of the penis. Sparsely crowded barnacles had more annulations in the penis and are inferred to have greater ability to stretch. At higher wave exposure, the diameter of the penis was greater, but the mass was not. This study identifies density of crowding as the most important cue that barnacles respond to when perceiving their mating group and details how penis morphology varies in response to wave exposure.     

Acquisition and loss of cold-tolerance in adult barnacles (Balanus balanoides) kept under laboratory conditions

In the laboratory, Balanus balanoides (L.) barnacles maintained without food at 5°C over the winter to summer period remained cold-tolerant. Winter animals maintained at 5°C and fed Artemia nauplii under a controlled light-dark regime until mid-summer lost this cold-tolerance. Summer animals, maintained without food at 5° or 15°C until midwinter, became cold-tolerant. Summer and winter animals subjected to increased (or decreased) ambient salinity for 48 h showed a small decrease (or increase) in their lower median lethal temperature (MLT—defined as the temperature at which 50% of the animals die after 18 h exposure to air). Summer animals subjected to desiccation for 48 h also showed a small decrease in their lower MLT. It is concluded that the seasonal changes in cold-tolerance of adult barnacles are probably induced by a combination of environmental factors including food availability, light intensity, day-length and changes in ambient sea-water temperatures. Development of cold-tolerance in the winter did not depend upon fertilisation. Changes in intra-cellular solute concentration which accompany adaptation to changes in ambient salinity or desiccation do not appear to be related to the seasonal changes in cold-tolerance.     

Effects of temperature and food availability on growth and reproduction in the neustonic pedunculate barnacle Lepas anserifera

To elucidate the life history of neustonic animals, growth and reproductive patterns were investigated in the hermaphroditic pedunculate barnacle Lepas anserifera in field and laboratory experiments in Wakayama, western Japan from 2006 to 2008. The effects of temperature (19, 24 or 29°C) and food availability (once or twice a week) on growth and reproduction were also studied in the laboratory. The barnacles grew and matured rapidly, especially in the field: individuals on the average grew from 3 mm to more than 12 mm in capitulum length within 15 days and some were brooding. High temperature and high food availability resulted in greater growth. High temperature also resulted in earlier maturation of both testes and ovaries, whereas the effect of food availability was less clear. The rapid growth and maturation, together with earlier maturation at higher temperatures, may be an adaptation to ephemeral floating objects to which they attach.     

大气沉降对土壤重金属累积的影响

选择大气污染严重的太原市为研究区,通过定点监测,分析了不同类型区大气TSP和降尘的污染状况及周年内的月变化趋势,测定了大气干湿沉降中Hg、Cd、Pb三种重金属元素的含量,并计算出大气干湿沉降对土壤系统重金属的年输入量分别为Hg4.48g/(hm2×a),Cd6.34g/(hm2×a),Pb349.4g/(hm2×a);不同类型区大气污染状况与大气沉降输入土壤中重金属累积顺序一致均为工矿区＞城区和近郊区＞风景区＞远郊区。     

Larval settlement in the echiuran worm Bonellia viridis: Settlement on both the adult proboscis and body trunk

It is widely documented that in Bonellia viridis Rolando (Echiura), the larvae settle on the proboscis of the adult female and differentiate into males. This study shows that although settlement on the proboscis is preferred, the larvae can also settle on the body trunk of the adult and differentiate into males. Aqueous extracts of homogenized trunk tissues are as effective as those of proboscis tissues in masculinizing the larvae. The natural secretion of the female body trunk, which is produced when the worm is disturbed, has potent masculinizing properties. The preferential settlement on the probiscis may not result from a different chemical nature of the substratum.     

Effects of salinity on fasted rainbow trout,Salmo gairdneri

Rainbow trout (Salmo gairdneri Richardson) which had been maintained for 120 days in salinities of fresh water, 7.5, 15.0 and 32.5 at 10°C were fasted for up to 48 days under these same environmental conditions. Live weight loss between Days 7 and 48 of starvation could be described by a straight line, as could the decrease in condition factor . Trout maintained in 32.5% S showed a significantly greater weight loss than those in salinities of 15.0 and below. Muscle water content increased slightly during fasting in fresh water, 7.5 and 15.0 S. In 32.5 S, however, muscle water fell significantly between Days 19 and 37. Liver water content also increased slightly during fasting, except in 32.5 S, where water content again decreased between Days 19 and 37. The volume of the gall bladder contents increased during fasting.     

Effects of salinity on multiplication and transmission of an intertidal trematode parasite

Salinity levels vary spatially in coastal areas, depending on proximity to freshwater sources, and may also be slowly decreasing as a result of anthropogenic climatic changes. The impact of salinity on host–parasite interactions is potentially a key regulator of transmission processes in intertidal areas, where trematodes are extremely common parasites of invertebrates and vertebrates. We investigated experimentally the effects of long-term exposure to decreased salinity levels on output of infective stages (cercariae) and their transmission success in the trematode Philophthalmus sp. This parasite uses the snail Zeacumantus subcarinatus as intermediate host, in which it asexually produces cercariae. After leaving the snail, cercariae encyst externally on hard substrates to await accidental ingestion by shorebirds, which serve as definitive hosts. We found that at reduced salinities (25 or 30 psu), the cercarial output of the parasite was lower, the time taken by cercariae to encyst was longer, fewer cercariae successfully encysted and encysted parasites had lower long-term survival than at normal seawater salinity (35 psu). The strong effect of salinity on the replication and transmission of this parasite suggests that there may be sources and sinks of transmission to birds along coastal areas, depending on local salinity conditions. Also, unless it evolves to adapt to changing conditions, the predicted reduction in salinity as a consequence of climate change may have negative impact on the parasite’s abundance.     

Effects of salinity fluctuation on the hemolymph composition of the blue crab Callinectes sapidus

The hemolymph of the blue crab Callinectes sapidus was hyperosmotic during 20-10-20 S and 30-10-30 S diurnal cycles. The hemolymph became isosmotic at 26 S and hyposmotic at 28 S in the 10-30-10 S diurnal cycle. Hemolymph Na⁺ was hyperionic to seawater throughout all cycles. Hemolymph Cl^- was hyperionic below 24 S and either isionic or hypoionic from 24 to 30 S. Hemolymph K⁺ concentrations were hyperionic below 26 S and either isionic or hypoionic from 26 to 30 S. Hemolymph Mg⁺⁺ values were hypoionic over the experimental salinity range (10 to 30). Hemolymph ninhydrin-positive substances (NPS) levels were directly related to ambient salinity.